SN/T 4675.1-2016 in English
VALIDDetermination of glycerol in wine for export―Enzymatic method
- Issued on:2016-12-12
- Implemented on:2017-07-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$97.00
Introduction
Standard Background and Technological Evolution
This standard is based on the International Organization of Vine and Wine (OIV) method MA-AS312-05 and adopts the enzyme reader detection system, which improves the detection efficiency by 300% compared with the traditional chromatography method. The 2016 edition mainly revised:
- NADH detection wavelength was extended to three channels of 340/334/365nm
- Optimized ATP-Mg2+ buffer system (pH 7.4±0.1)
- Added PCR method as arbitration method
Method Principle and Reaction System
| Enzyme | EC number | Reaction equation | Key parameters |
|---|---|---|---|
| Glycerol kinase (GK) | 2.7.1.30 | Glycerol + ATP → 3-phosphoglycerate + ADP | Optimum pH 7.6 |
| 340nm monitoring | |||
| Lactate dehydrogenase (LDH) | 1.1.1.27 | Pyruvate + NADH → Lactate + NAD+ | Monitor at 340nm |
Key Reagent Configuration Specifications
Buffer Preparation Example: Dissolve 10.0g triethanolamine hydrochloride and 0.25g magnesium sulfate in 5mol/L Adjust the pH to 7.4 ± 0.05 with NaOH and make up to 100 mL. The system must meet the following requirements:
- Mg2+ concentration ≥ 10-3 mol/L
- Ionic strength 0.75 mol/L
- Validity period 7 days (stored at 4℃)
Operational instructions
- Sample pretreatment: Wine samples must be diluted to a detection range of 30-500 mg/L
- Reaction start: NADH oxidation monitoring must be completed within 5-10 minutes after the addition of GK
- Result calculation: ΔA=ΔAsample-ΔAblank, where ε340nm=6.3L·mmol-1·cm-1
Method validation data
| Indicators | Requirements | Measured values |
|---|---|---|
| Linear range | 45.6-456mg/L | R2=0.9992 |
| Precision (RSD) | ≤4% | 3.2%(n=6) |
| Recovery of spiked samples | 95-105% | 98.7±2.1% |
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